Post-transcriptional modification of nucleic acids including transfer RNA (tRNA), ribosomal RNA (rRNA) and messenger RNA (mRNA) is vital for fine-tunning of mRNA translation. Methylation is one of the most widespread post-transcriptional modifications in both eukaryotes and prokaryotes. HsWBSCR22 and ScBUD23 encodes a 18S rRNA methyltransferase that positively regulates cell growth by mediating ribosome maturation in human and yeast, respectively. However, presence and function of 18S rRNA methyltransferase in green algae are still elusive. Here, through bioinformatic analysis, we identified CrBUD23 as the human WBSCR22 homolog in genome of the green algae model organism Chlamydonomas reinhardtii. CrBUD23 was a conserved putative 18S rRNA methyltransferase widely exited in algae, plants, insects and mammalians. Transcription of CrBUD23 was upregulated by high light and down-regulated by low light, indicating its role in photosynthesis and energy metabolism. To characterize its biological function, coding sequence of CrBUD23 fused with a green fluorescence protein (GFP) tag was derived by 35S promoter and stably integrated into Chlamydomonas genome by glass bead-mediated transformation. Compared to C. reinhardtii wild type CC-5325, transgenic strains overexpressing CrBUD23 resulted in accelerated cell growth, thereby leading to elevated biomass, dry weight and protein content. Moreover, overexpression of CrBUD23 increased content of photosynthetic pigments but not elicit the activation of antioxidative enzymes, suggesting CrBUD23 favors growth and proliferation in the trade-off with stress responses. Bioinformatic analysis revealed the G1177 was the putative methylation site in 18S rRNA of C. reinhardtii CC-849. G1177 was conserved in other Chlamydonomas isolates, indicating the conserved methyltransferase activity of BUD23 proteins. In addition, CrTrm122, the homolog of BUD23 interactor Trm112, was found involved in responses to high light as same as CrBUD23. Taken together, our study revealed that cell growth, protein content and lutein accumulation of Chlamydomonas were positively regulated by the 18S rRNA methyltransferase CrBUD23, which could serve as a promising candidate for microalgae genetic engineering.
Microalgal biotechnology has become a promising field of research for the production of valuable, sustainable and environmentally friendly byproducts, especially for carotenoids. Bulk accumulation of secondary carotenoids in microalgae are mostly induced by oxidative stress of cells. In this research, we investigated the effects of antioxidant enzyme activity inhibition on carotenogenesis in a microalga Dactylococcus dissociatus MT1. The activities of four major antioxidant enzyme families, namely superoxide dismutase (SOD), catalases (CAT), glutathione peroxydases (GPX) and ascorbate perxodases (APX), were inhibited by relevant inhibitors during the stressed cultivation of D. dissociatus to observe the effects on carotenogensis. A 91% decrease in activity was observed for CAT, comparing with controls without any inhibitors added, followed by 65%, 61%, and 47% for the enzymes SOD, APX, and GPX, respectively. Concomitantly, it was found that this partial inhibition had substantial influences on the accumulation of carotenoids, with the highest production levels obtained in CAT inhibition conditions and an increase of 2.6 times of carotenoid concentration observed, comparing with control cultivation conditions. We conclude that the modulation of antioxidant enzyme activities could lead to the overproduction of carotenoids in this microalgal cell culture, and we expect that this novel approach of optimizing carotenogenesis processes for D. dissociatus cell cultures could be transferrable to other cell culture systems and might have an important impact on the carotenoid production industry.
Frontiers in Bioengineering and Biotechnology
Versatility of Algae in Addressing the Global Sustainability Challenges